Objectlve To explore the mechanism of the protection in high expression of HO-1 induced by CoPP on murine islet xenografts. Method An islet transplantation of a SD rat-to-C57 BL/6 mouse model was established.Mice were randomized into five groups i.e.control,CoPP-induction in vivo,CoPP+ZnPP in vivo.CoPP-induction in vitro and CoPP+ZnPP in vitro and the islet xenografts were transplanted into the subrenal capsule.Normoglycemia time was recorded and insulin-releasing test was performed.IL-10、TNF-α、IL-1β and INF-γ in serum and their cytokine mRNA and HO-1 in xenografts were measured by RT-PCR and Western-blotting.The pathological examination was done to observe the lymphocyte infiltration. Results There Was significant difference in the normoglycemia time between CoPP-induction in vivo and in vitro and other three groups.The results of insulin-releasing stimulated by low level glucose were identical among groups,but that of insulin-releasing stimulated by high-glucose in in vivo group were the hiishest as in CoPP-induction in vivo and in vitro and control group were 187.68 ±19.93、137.22±11.73,91.25±12.64 μIU·ml~(-1)·10islets~(-1)·45 min~(-1),(P<0.05).The IL-10 in serum in CoPP-induction in vivo and in vitro(in vivo:72.97±9.74 pg/ml;in vitro:70.84±3.56 pg/ml)was significantly hisher than other three groups(control:30.57±3.93 pg/ml;CoPP+ZnPP in vivo:39.78±3.00 pg/ml;CoPP+ZnPP in vitro:35.42±4.30 pg/ml).The expression tendency of IL-10 mRNA was similar to that of insulin secretion.There was no significant difieFence in TNF-α、IL-1β and INF-γ.The expression of HO-1 by PCR and Western-blot analysis in CoPP-induction in vivo and vitro was higher than other three groups.The pathological examination showed that fewer lymphocytes infiltrated into the islet xenografts from CoPP-treated in comparison with xenografts from other three groups. Conclusion HO-1 could improve the survival of islet xenograft:the induetion of HO-1 expression in vivo was much mole efficient than in vitro.The CoPP-induction could be related to immune modulation of IL-10.

Objective To analyze the dose-effect relationship between cobalt protoporphyrin (CoPP) and heme oxygenase-1 (HO-1) expression in islets and to investigate the protective effect of strongly expression of HO-1 in islet xenotransplantation. Methods Donor islets isolated and purified from SD rats were randomly divided into 5 groups and incubated with different doses of CoPP for 24 h.Group A: 0 mmol/L; Group B: 5 mmol/L; Group C: 25 mmol/L; Group D: 50 mmol/L; Group E:75 mmol/L. The expression of HO-1 mRNA and protein in islets was detected by RT-PCR and Western blot respectively. Glucose of low and high concentrations was added to islets in vitro to test insulin-releasing function. The optimal dose of CoPP which could induce the strongest HO-1 expression was chosen according to the results. Recipients were randomly divided into 2 groups. Control group received untreated xeno-islets, and the experimental group received islets incubated with optimal CoPP close in vitro. Glycemia and rejection were observed after transplantation daily. Results The expression of HO-1 mRNA and protein in xeno-islets of group D was significantly higher than in other groups (P＜0.05). After stimulation of glucose, the insulin concentration in group D was significantly higher than in other groups (P＜0.05). The optimal dose of CoPP which could induce the strongest HO-1 expression was 50 mmol/L. The time for normoglyeemia in experimental group was (14.63±1.19) days, significantly longer than that in control group (9.88±2.17)days (P＜0.01). Conclusion The strongest expression of HO-1 induced by CoPP in vitro promotes the glucose-stimulated insulin secretion of islets and prolonged the survival of xeno-islet grafts by protecting them from rejection.

Objective To investigate the effects of peritoneal injection of cobaltic protoporphyrin Ⅸ chloride(CoPP)to induce heme oxygenase-1(Ho-1)upregulation in rat islet cells.Mthods Forty rats were divided into 5 groups by management 24 h before islets isolation:group A received inlzaporitoneal injection with 2.5 ms/ks CoPP,group B with 5 ms/ks CoPP,group C with 7.5 ms/kg CoPP,and group D with 10 mg/kg CoPP.In control group NS was used instead.A modified Goth approach was used for islet isolation.the yield and purity of the islets were assessed.The expression of Ho-1 mRNA and protein were detected by real time-PCR and Western blotting respectively.Islet function was tested by glucose stimulation test.Result Severe damage was found in tlle rats in group C and D.There was no difference in islet yield and purity for group A、B、C and control(P＞0.05).Group B had the highest Ho-1 mRNA and protein expression among the 4 groups.Though there was no difference in insulin secretion by low glucose challenge for group A、B and control's islets(P＞0.05),when challenged by hish level of glucose,significant deviation was observed.The imulin secretion level was(172.37±16.4)、(187.68±19.93)and(91.25±Conclusion Peritoneal iajection of 5 mg/kg CoPP can significantly enhance the expression of Ho-1 mRNA and protein in tat islet safely and enhance the function of islet when challenged by hish concentration of glucose.

BACKGROUNDAtrial fibrillation (AF) is one of the most common arrhythmia after coronary artery bypass grafting (CABG), which not only increases the suffering of the patients, but also prolongs hospital stay and enhances cost of care, especially for patients older than 70 years. This study was designed to evaluate the efficacy and safety of low-dose amiodarone in the prevention of AF after CABG, especially for the elderly.

METHODSTwo hundred and ten senile patients undergoing off-pump CABG were included in this prospective, randomized, double-blind and placebo controlled study. Patients were given 10 mg/kg of amiodarone (low-dose amiodarone group, n = 100) or placebo (control group, n = 110) daily for 7 days before surgery and followed by 200 mg of amiodarone or placebo daily for 10 days postoperatively.

RESULTSPostoperative AF occurred in 16 patients (16%) receiving amiodarone and in 36 (37.7%) patients receiving placebo (P = 0.006). AF occurred at (58.13 +/- 16.63) hours after CABG in the low-dose amiodarone group and at (45.03 +/- 17.40) hours in the control group (P = 0.018). The maximum ventricular rate during AF was significantly slower in the low-dose amiodarone group ((121.42 +/- 28.91) beats/min) than in the control group ((134.11 +/- 30.57) beats/min, P = 0.036). The duration of AF was (10.92 +/- 9.56) hours for the low-dose amiodarone group compared with (14.81 +/- 10.37) hours for the control group (P = 0.002). The postoperative left ventricular ejection fraction (LVEF) was significantly improved in the low-dose amiodarone group (from (59.9 +/- 10.3)% to (63.4 +/- 11.4)%, P = 0.001), and significantly higher compared with the control group ((58.5 +/- 10.7)%, P = 0.002). Both groups had a similar incidence of complication other than rhythm disturbances (12.0% vs 16.4%, P = 0.368). The low-dose amiodarone group patients had shorter hospital stays ((11.8 +/- 3.2) days vs (13.8 +/- 4.7) days, P = 0.001) and lower cost of care (RMB (79 115 +/- 16 673) Yuan vs RMB (84 997 +/- 21 587) Yuan, P = 0.031) than that of control group patients. The in-hospital mortality was not significantly different between the two groups (1.0% vs 0.9%, P = 0.946).

CONCLUSIONSPerioperative low-dose oral amiodarone appeared to be cost-effective in the prevention and delay of new-onset postoperative AF in aged patients. It significantly reduced ventricular rate and duration of AF after CABG, decreased hospital cost and stay, as well as promoted the amelioration of left ventricular systolic function. Furthermore, low-dose amiodarone was safe to use and well tolerated with low toxic and side effects, and did not increase the risk of complications and mortality. It is proved to be a first-line therapy and as routine prophylaxis for AF after CABG, especially for elderly patients complicated with left ventricular dysfunction.

Aged ; Amiodarone ; administration & dosage ; Anti-Arrhythmia Agents ; administration & dosage ; Atrial Fibrillation ; etiology ; prevention & control ; Coronary Artery Bypass ; adverse effects ; Double-Blind Method ; Drug Administration Schedule ; Female ; Humans ; Male ; Treatment Outcome

A quantitative HPLC-DAD method was developed for simultaneous determination of N-trans-p-coumaroyloctopamine and N-trans-p-coumaroyltyramine in Solani Melongenae Radix from different cultivation regions in China The separation was performed on an Agilent Eclipse XDB C18 column (4.6 mm x 250 mm, 5 microm) at 30 degrees C with a gradient elution of methanol and 0.1% formic acid in water as mobile phase. The flow rate was set at 1.0 mL x min(-1) and the detection wavelength was 300 nm. The calibration curves of N-trans-p-coumaroyloctopamine and N-trans-p-coumaroyltyramine were linear over the ranges of 2.84-68.16, 3.10-74.40 mg x L(-1), and the average recoveries (n = 9) were 99.30% and 102.8%, respectively. The developed method was successfully applied for the analysis of sixteen samples from different cultivation regions in China, which indicated that the method is simple, rapid, accurate, and reliable for quality evaluation of Solani Melongenae Radix.
Amides ; analysis ; China ; Chromatography, High Pressure Liquid ; methods ; Drugs, Chinese Herbal ; analysis ; Plant Roots ; chemistry ; Solanaceae ; chemistry ; classification

OBJECTIVETo establish criteria for AIS of Lenke5 and Lenke6 by an anterior only procedure of the lower curve fusion.

METHODSA retrospective study was conducted between March 1999 and May 2004 to investigate 52 AIS patients of Lenke5 and Lenke6. All the patients were observed 24 years (34 months on average). Many parameters were evaluated. At final assessment, two groups emerged: Group A had satisfactory results (the thoracic curve was reduced) and Group B had just the opposite.

RESULTSPreoperative thoracic curve in group A averaged 33 degrees and 18 degrees after surgery. The lumbar curve averaged 49 degrees before surgery and 21 degrees after surgery. In group B (n = 6), the average thoracic curve was 38 degrees before surgery and 45 degrees after surgery, whereas the lumbar curve averaged 46 degrees before surgery and 25 degrees after surgery. Two of these patients underwent posterior thoracic instrumentation and fusion because of the unreasonable balance.

CONCLUSIONSA successful surgical outcome was dependent on both the flexibility of the thoracic curve and the patients' maturity. The thoracolumbar/lumbar-thoracic (TL/L:T) Cobb ratio in combination with the flexibility of the thoracic curve were the best predictors among the structural indexes.

Adolescent ; Adult ; Child ; Female ; Follow-Up Studies ; Humans ; Male ; Retrospective Studies ; Scoliosis ; surgery ; Spinal Fusion ; methods ; Treatment Outcome

In order to elucidate the effect of acetylcholine (ACh) on the occurrence and development of human pituitary adenoma, it was firstly observed whether there exists choline acetyl transferase (ChAT) which is necessary for the synthesis of acetylcholine in the cells of human pituitary adenoma, and then MTT method, (3)H TdR incorporation, cell cycle analysis and TUNEL were employed to estimate the influence of ACh on the proliferation, DNA synthesis and apoptosis of three kinds of human pituitary adenoma (human prolactinoma, somatotropinoma and non-functional tumor) cells cultured in vitro. The results showed that (1) the positive staining of ChAT was obviously observed in the cells of the three kinds of human pituitary adenoma, however, it was lower than that in normal human pituitary gland; (2) ACh had a similar effect on the proliferation of the three kinds of human pituitary adenoma cells. ACh at 0.1-10 micromol/L decreased the (3)H TdR incorporation and the MTT A value in a dose-dependent manner. At the same time, ACh decreased the ratio of S or G(2) phase pituitary adenoma cells significantly, but increased the ratio of G(1) phase pituitary tumour cells markedly; (3) the effect of acetylcholine on the proliferation of human pituitary adenoma cells was inhibited by atropine, but not by tubocurarine; (4) ACh had no effect on the apoptosis of human pituitary adenoma cells cultured in vitro. These data suggest that ACh may have a significant modulating effect on the proliferation of pituitary adenoma cells by means of paracrine or autocrine, and the effect is mediated by muscarinic receptor.
Acetylcholine ; pharmacology ; physiology ; Acetyltransferases ; biosynthesis ; physiology ; Adenoma ; pathology ; secretion ; Apoptosis ; drug effects ; Cell Division ; drug effects ; Dose-Response Relationship, Drug ; Humans ; Pituitary Neoplasms ; pathology ; secretion ; Tumor Cells, Cultured

BACKGROUNDThe induced expression of heme oxygenase-1 (HO-1) in donor islets improves allograft survival. Cobalt protoporphyrin (CoPP) could significantly enhance the expression of HO-1 mRNA and protein in rat islet safely. Our work was to study how to protect pancreatic islet xenograft by CoPP-induction.

METHODSIslet xenografts treated with CoPP-induction and CoPP + Zinc protoporphyrin (ZnPP) in vitro and in vivo were randomly transplanted into murine subrenal capsule; then the graft survival time was compared by blood glucose level and pathological examination and meanwhile the interferon γ (IFN-γ), tumor necrosis factor α (TNF-α), interleukin 10 (IL-10) and IL-1β level in serum and their mRNA and HO-1 mRNA and protein expression were examined.

RESULTSIslets with CoPP-induction under low- and high-glucose stimulation exhibited much higher insulin secretion compared with other three groups. CoPP-induction could increase higher expression of HO-1 (mRNA: 3.33- and 76.09-fold in vitro and in vivo; protein: 2.85- and 58.72-fold). The normoglycemia time in induction groups ((14.63 ± 1.19) and (16.88 ± 1.64) days) was significantly longer. The pathological examination showed less lymphocyte infiltration in induction groups. The IL-10 level and its mRNA in induction groups were significantly higher.

CONCLUSIONSThe HO-1 induced by CoPP would significantly improve function, prolong normoglycemia time and reduce lymphocyte infiltration. Meanwhile CoPP-induction in vivo had more beneficial effects than in vitro. Its mechanism could be related to immune-modulation of IL-10.

Animals ; Blotting, Western ; Graft Survival ; Heme Oxygenase-1 ; genetics ; metabolism ; Interleukin-10 ; blood ; Islets of Langerhans ; enzymology ; metabolism ; Islets of Langerhans Transplantation ; Male ; Mice ; Mice, Inbred BALB C ; Rats ; Rats, Sprague-Dawley ; Reverse Transcriptase Polymerase Chain Reaction ; Transplantation, Heterologous

OBJECTIVETo study the protective effect of islet xenograft and its possible mechanism of high expression of heme oxygenase-1 (HO-1) in donor pancreas islet induced by cobalt protoporphyrin (CoPP).

METHODSMale SD rats and C57BL/6 mouse were used as donors and recipients respectively. Donors were divided into 3 groups according to different pretreatment 24 hours before donation: control group (injected intraperitoneally with NaCl), induce group [injected intraperitoneally with cobalt-protoporphyrin (CoPP)], block group (injected intraperitoneally with CoPP and zinc protoporphyrin simultaneously). A modified approach was used for islet isolation.Recipients were rendered diabetic by intraperitoneal injection of streptozotocin. Islets were transplanted into mouse subrenal capsule. Postoperative mouse glycemia were monitored daily and normoglycemia time was compared among each group. The receptor mouse serum IL-10 was detected by ELISA approach, and real-time PCR was used to check the expression of IL-10 mRNA in islet graft tissues. The graft tissues were observed for the lymphocyte infiltration after HE staining.

RESULTSDiabetes mice accepted islets untreated, induced or blocked maintained the euglycemia for (9.3 +/- 1.4), (16.3 +/- 1.5) and (9.7 +/- 1.0) d respectively. The xeno-islets presented HO-1 over-expression survived much longer than that absent (P < 0.05), it was no significance between control group and block group (P > 0.05). The mouse islet serum IL-10 content after induction was (73.0 +/- 9.7) pg/ml, significantly higher than (30.6 +/- 3.9) pg/ml of the untreated group and (32.1 +/- 5.9) pg/ml of the blocked group (P < 0.05), there was no difference between control group and block group (P > 0.05). Moreover, the IL-10 mRNA expression up-regulated statistic significantly in HO-1 induced islet xeno-graft. Pathological examination showed that the graft lymphocyte infiltration of the induced group was obviously less serious than the other two groups.

CONCLUSIONSThe higher expression of HO-1 induced by CoPP in vivo would significantly prolong graft survival time and its mechanism could be related to immune modulation of IL-10.

Animals ; Diabetes Mellitus, Experimental ; metabolism ; pathology ; surgery ; Graft Survival ; Heme Oxygenase-1 ; drug effects ; metabolism ; Interleukin-10 ; metabolism ; Islets of Langerhans ; metabolism ; pathology ; Male ; Mice ; Mice, Inbred C57BL ; Pancreas Transplantation ; Protoporphyrins ; pharmacology ; Rats ; Rats, Sprague-Dawley ; Subrenal Capsule Assay ; Transplantation, Heterologous

Objective: To explore the relevant gene, signaling pathway for permanent atrial fibrillation (pAF) occurrence in order to provide the molecular basis of the pathogenesis of pAF. Methods: Our research included in 2 groups: pAF group, n=7 patients and Control group, n=4 healthy subjects with sinus rhythm. Agilent 4x44K microarray was used to analyze the mRNA in left atrium for differential gene expression profile. Based on Gene Ontology, KEGG and Biocarta databases, differentially expressed genes were studied for their relevant function and signaling pathway. Furthermore, the genes with significant differences were verified by quantitative real time PCR (qRT-PCR) in pathological specimen from 5 pAF patients and 5 normal heart donors. Results: The expression profile identified 987 abnormally expressed genes, 567 of them were down-regulated and 420 were up-regulated. 9 genes with significant differences were verified by qRT-PCR in pathological specimen and the changes were similar to microarray; those genes were closely related to pAF by involving left atrium fibrosis, electrical remodeling, inflammation, cellular stress response, metabolism and transcription regulation. GO and Pathway analysis indicated that down-regulated genes were mainly involved in metabolic processes; up-regulated genes had the effects on cellular stress response, immune response and platelet activation. Conclusion: Microarray technology identified some important genes related to pAF occurrence; such genes involved in left atrial structural and functional remodeling via affecting cellular metabolism, inflammation, immune response and thrombogenesis in relevant patients.